Effects of Pb and CO2 on Soil Microbial Community Associated with Pinus densiflora-Lab

소나무(Pinus densiflora) 생육토양의 미생물 군집에 미치는 납과 CO2의 영향

  • Hong, Sun-Hwa (Department of Environmental Science and Engineering, Ewha Woman's University) ;
  • Kim, Sung-Hyun (Department of Life Science, Ewha Woman's University) ;
  • Kang, Ho-Jeong (Department of Environmental Science and Engineering, Ewha Woman's University) ;
  • Ryu, Hee-Wook (Department of Chemical and Environmental Engineering, Soongsil University) ;
  • Lee, Sang-Don (Department of Environmental Science and Engineering, Ewha Woman's University) ;
  • Lee, In-Sook (Department of Life Science, Ewha Woman's University) ;
  • Cho, Kyung-Suk (Department of Environmental Science and Engineering, Ewha Woman's University)
  • 홍선화 (이화여자대학교 환경공학과) ;
  • 김성현 (이화여자대학교 생명과학과) ;
  • 강호정 (이화여자대학교 환경공학과) ;
  • 류희욱 (숭실대학교 환경화학공학과) ;
  • 이상돈 (이화여자대학교 환경공학과) ;
  • 이인숙 (이화여자대학교 생명과학과) ;
  • 조경숙 (이화여자대학교 환경공학과)
  • Published : 2006.12.31


Effects of Pb and $CO_2$ on soil microbial community associated with Pinus densiflora were investigated using community level physiological profiles (CLPP) and 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) methods. Two-years pine trees were planted in Pb-contaminated soils and uncontaminated soils, and cultivated for 3 months in the growth chamber where $CO_2$ concentration was controlled at 380 or 760 ppmv. The structure of microbial community was analyzed in 6 kinds of soil samples (CA-0M : $CO_2$ 380 ppmv + Pb 0 mg/kg + initial, CB-0M : $CO_2$ 380 ppmv + Pb 500 mg/kg + initial, CA-3M : $CO_2$ 380 ppmv + Pb 0 mg/kg + after 3 months, CB-3M : $CO_2$ 380 ppmv + Pb 500 mglkg + after 3 months, EA-3M : $CO_2$ 760 ppmv + Pb 0 mg/kg + after 3 months, EB-3M : $CO_2$ 760 ppmv + Pb 500 mg/kg + after 3 months). After 3 months, the substrate utilization in the uncontaminated soil samples (CA-3M vs EA-3M) was not significantly influenced by $CO_2$ concentrations. However, the substrate utilization in the Pb-contaminated soil samples (CB-3M vs EB-3M) was enhanced by the elevated $CO_2$ concentrations. The results of principal component analysis based on substrate utilization activities showed that the structure of microbial community structure in each soil sample was grouped by Pb-contamination. The similarities of DGGE fingerprints were 56.3 % between the uncontaminated soil samples (CA-3M vs EA-3M), and 71.4% between the Pb-contaminated soil samples (CB-3M vs. EB-3M). The similarities between the soil samples under $CO_2$ 380 ppmv (CA-3M vs CB-3M) and $CO_2$, 760 ppmv (EA-3M vs EB-3M) were 53.3% and 35.8%, respectively. These results suggested that the structure of microbial community associated with Pinus densiflora were sensitively specialized by Pb-contamination rather than $CO_2$ concentration.


  1. Amann RI, Ludwig W, Schleifer KH. 1995. Phylogenetic identification and in situ detection of individual mmicrobial cells without cultivation. Microbial Rev 59: 143-169
  2. Bruns MA, Stephen JR, Kowalchuk GA, Prosser JI, Paul EA. 1999 Comparative diversity of ammonia oxidizer 16S-rRNA gene sequences in native, tilled, and successional soils. Appl Envrion Microbiol 65: 2994-3000
  3. Cho WS, Lee EH, Shim EH, Kim JS, Ryu HW, Cho KS. 2005. Bacterial communities of biofilm sampled from seepage groundwater contaminated with petroleum oil. J Microbiol Biotechnol 15: 952-964
  4. Christiane M, Morten M, Heribert I. 1999. Elevated $CO_{2}$ alters community- level physiological profiles and enzyme activites in alpine grassland. J Microbiol Methods 36: 35-43
  5. Duineveld BM, Rosado AS, Elsas JD, Veen JA. 1998. Analysis of the dynamics of bacterial communities in the rhizosphere of the chrysanthemum via denaturing gradient gel electrophoresis and substrate utilization pattern. Appl Envrion Microbiol 64: 4950-4957
  6. Head IM, Saunders JR, Pickup RW. 1998. Microbial evolution diversity, and ecology: a decade of ribosomal RNA analysis of uncultivated microorganisms. Microbial Ecol 35: 1-21
  7. Hershey DR. 1994. Solution culture hydroponics: History and inexpensive equipment. American Biology Teacher 56: 111-118
  8. Heuer HM, Baker KP, Smalla K, Wellington EMH. 1997. Analysis of actinomycete communities by specific amplification of genes encoding 16S-rRNA and gel-electrophoretic separation in denaturing gradients. Appl Envrion Microbiol 63: 3232-3241
  9. Garland JL, Aaron LM. 1991. Classification and characterization of heterotrophic microbial communities on the basis of communitylevel sole-carbon-source utilization. Appl Envrion Microbiol 57: 2351-2359
  10. Giovannoni SJ, Brischgi TB, Moyer CL, Field KS. 1990. Genetic diversity in sargasso sea bacterioplankton. Nature (London) 345: 60-63
  11. Glick BR, Karaturovic DM, Newell PC. 1995. A Novel procedure for rapid isolation of plant growth promoting Pseudomonads. Can J Microbiol 41: 533-536
  12. Glick BR. 2003. Phytoremediation : Synergistic use of plants and bacteria to clean up the environment. Biotechnol Adv 21: 383-393
  13. Kennedy IR, Pereg-Gerk LL, Wood C, Deaker R, Gilchrist K, Katupitiya S. 1997. Biological nitrogen fixation in non-leguminous field crops: Facilitating the evolution of an effective association between azospirillum and wheat. Plant Soil 194: 65-79
  14. Kloepper JW, Lifshitz R, Zablotowicz RM. 1989. Freeliving bacterial iinocula for enhancing crop productivity. Trends Biotechnol 7: 39- 44
  15. Knight B, McGrath SP, Chaudri AM. 1997. Biomass carbon measurements and substrate utilization patterns of microbial populations from soils amended with cadmium, copper, or zinc. Appl Environ Microbiol 63: 39-43
  16. Ian FS, Peter JF. 2003. A tribute to claude shannon (1916-2001) and a plea for more rigorous use of species richness, species diversity and the shannon-wiener index. Global Ecol Biogeog 12: 177-179
  17. Lori RJ, Nicholas LA, Jhon M, Steven TR, Nancy CT, Jhon JK. 2005. Elevated atmospheric $CO_{2}$ alters soil microbial communities associated with trembling aspen (Populus tremuloides) roots. Microbial Ecol 50: 102-109
  18. Marcus S, Ueli AH, Georeg RH, Michael JS. 1996. Microbial community Changes in the rhizospheres of white clover and perennial ryegress exposed to free air carbon dioxide enrichment (FACE). Soil Biol Biochem 28: 1717-1724
  19. Matthias CR, Kate MS, Jhon NK. Michael FA. 1997. Microbial Carbonsubstrate utilization in the rhizosphere of Gutierrezia sarothrae grown in elevated atmospheric carbon dioxide. Soil Biol Biochem 29: 1387-1394
  20. Mordukhova EA, Skvortsova NP, Kochetkov VV, Dubeikovskii AN, Boronin AM. 1991. Synthesis of the phytohormone indole-3-Acetic acid by rhizosphere bacteria of the genus Pseudomonas. Mikrobiologiya 60: 494-500
  21. Muhammad A, Xu J, Li Z, Wang H, Yao H. 2005. Effects of lead and cadmium nitrate on biomass and substrate utilization pattern of soil microbial communities. Chemosphere 60: 508-514
  22. Muyzer G, De-Waal EC, Uitterlinden AG. 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59: 695-
  23. Nicola L, Therese H, Tanja K, Arata K, Tsuyoshi Y, Petra M, Ellen K. 2006. Response of microbial activity and microbial community composition in soil to long-term arsenic and cadmium exposure. Soil Biol Biochem (in press)
  24. Shanahan P, O'Sullivan DJ, Simpson P, Glennon JD, O'Gara F. 1992. Isolation of 2,4-diacetylphloroglucinol from a fluorescent Pseudomonad and investigation of physiological parameters influencing its production. Appl Environ Microbiol 58: 353-358
  25. Pearce D, Bzin MJ, Lynch JM. 1995. The rhizosphere as a biofilm. In: Lappin-Scott, H. M., Costerton, J. W. (Eds), Microbial Biofilms. cambridge University Press, Cambridge. 207-220
  26. Ward DM, Weller R, Bateson MM. 1990. 16s rRNA Sequences reveal numerous uncultured microorganisms in a natural community. Nature 345: 63-65
  27. Yang X, Feng Y, He Z, Stoffella PJ. 2005. Molecular mechanisms of heavy metal hyperaccumulation and phytoremediation. J Trace Elem Med Biol 18: 339-353
  28. Yang Y, Campell CD, Clark L, Cameron CM, Paterson E. 2006. Microbial indicators of heavy metal contamination in urban and rural Soils. Chemosphere 11:1942-52
  29. Yao H, Xu J, Huang C, Campell CD. 2000. Microbial biomass and community structure in a accumulation in soils increasing fertility and changing land use. Microbial Ecol 40: 223-237
  30. Yao H, Xu J, Huang C. 2003. Structure utilisation pattern, biomass and activity of microbial communities in a sequence of heavy metal polluted paddy soils. Geoderma 115: 139-1487